Toxicological Sciences最新文献

{"title":"Culture media influences primary human bronchial epithelial cell morphology, differentiation status, and transcriptional response to ozone exposure.","authors":"Sarah A Lester, Sabri H Abdelwahab, Scott H Randell, Samir N P Kelada","doi":"10.1093/toxsci/kfaf089","DOIUrl":"10.1093/toxsci/kfaf089","url":null,"abstract":"<p><p>Exposure to the ambient air pollutant ozone induces acute and chronic respiratory health effects in part by causing inflammation of the airways. Several aspects of the inflammatory response to ozone can be modeled in vitro using primary human bronchial epithelial cells (HBECs) cultured at an air-liquid interface. We tested two commonly used HBEC culture media systems, one proprietary and one non-proprietary, to identify which system yielded the most in vivo-like pro-inflammatory response to acute ozone exposure as reflected by gene expression. Cells from 6 donors were grown in each culture system in parallel, followed by examination of epithelial morphology and cell type proportions prior to ozone exposure. Cultures grown in the proprietary system were notably thicker and contained more ciliated and secretory cells, as well as internal cyst-like structures. The transcriptomic response to acute ozone exposure (0.5 parts per million ozone × 2 h) was strongly affected by media type. HBECs grown in the proprietary system exhibited minimal changes after ozone, with only 7 differentially expressed genes (DEGs). In contrast, HBECs grown in the non-proprietary system exhibited a more dynamic response with 128 DEGs, including hallmark response genes indicative of inflammation (CXCL8) and oxidative stress (HMOX1). Gene set enrichment analysis using the 128 DEGs further corroborated upregulation of oxidative stress and inflammation pathways. In total, our results indicate that the choice of HBEC culture media should be carefully considered to best model the in vivo response to ozone.</p>","PeriodicalId":23178,"journal":{"name":"Toxicological Sciences","volume":" ","pages":"31-36"},"PeriodicalIF":4.1,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12448227/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144699630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of four mechanisms of toxicity for per- and polyfluoroalkyl substances through transcriptomic profiling in human liver spheroids exposed to 24 PFAS.","authors":"Greggory C Addicks, Andrea Rowan-Carroll, Karen Leingartner, Andrew Williams, Mathew J Meier, Luigi Lorusso, Carole L Yauk, Ella Atlas","doi":"10.1093/toxsci/kfaf075","DOIUrl":"10.1093/toxsci/kfaf075","url":null,"abstract":"<p><p>Per- and polyfluoroalkyl substances (PFAS) are persistent and widespread contaminants. Epidemiological effects of PFAS include increased serum cholesterol, decreased immune response to vaccination and disease, and increased incidence of cancer; however, PFAS modes of action remain unclear. Herein, we analyzed gene expression data from human liver spheroids that were exposed to several concentrations of 24 different PFAS. Benchmark concentration (BMC) response modeling was used to identify the 250 lowest gene BMCs for each PFAS. Hierarchical clustering analysis revealed 4 functionally diverse gene sets. Each gene set was affected by a distinct group of PFAS, whereas individual PFAS were usually part of more than 1 PFAS group. The biological roles of these gene sets relate to: (1) cholesterol biogenesis and cholesterol clearance (downregulated by 7 fluorocarbon or longer PFAS), putatively through discordance of cholesterol sensing by SCAP and LXR due to membrane integration of PFAS; (2) lipolysis (upregulated by 8 carbon or shorter PFAS); (3) innate immunity (downregulated by most PFAS); and (4) adaptive immunity (downregulated by sulfonate-type PFAS). The distinctions between the 4 PFAS groups suggest that PFAS can act through at least 4 independent mechanisms. The molecular characteristics of each PFAS group may be useful for understanding the molecular interactions leading to their effect on gene expression. Inclusion of some PFAS congeners in more than one PFAS group suggests that individual PFAS can act through multiple unrelated molecular interactions. This transcriptomic analysis offers a major advancement to the understanding of the molecular mechanisms underlying the effects of PFAS exposure and provides guidance for future work that may strengthen links between PFAS exposure and their proposed effects on human health.</p>","PeriodicalId":23178,"journal":{"name":"Toxicological Sciences","volume":" ","pages":"161-180"},"PeriodicalIF":4.1,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144275957","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Acetate derived from metabolism of ethanol affects gene expression in bone and contributes to delays in chondrogenic differentiation.","authors":"Kim B Pedersen, Cheyleann Del Valle Ponce de Leon, Hardy Hang, Jin-Ran Chen, Christopher E Randolph, Jovanny Zabaleta, Christopher M Taylor, Meng Luo, Alexandra Denys, Martin J J Ronis","doi":"10.1093/toxsci/kfaf073","DOIUrl":"10.1093/toxsci/kfaf073","url":null,"abstract":"<p><p>Alcohol intake is a risk factor for the development of osteopenia. Ethanol perturbs gene expression in osteoblasts and osteoclasts and disrupts growth plate morphology. Hepatic metabolism of ethanol to acetate elevates concentrations of acetate in the circulation. We investigated whether acetate could, in part, mediate the toxicity of ethanol in bone and on chondrocyte differentiation. When ethanol and acetate were compared by gavage for 4 consecutive days, none of 11 selected genes involved in bone homeostasis were significantly affected by acetate, but acetate responses significantly correlated with ethanol responses. Intraperitoneal injection with acetate to transiently elevate serum acetate for 4 consecutive days significantly increased expression of 2 markers of osteoclast differentiation, calcitonin receptor (Calcr) and Ocstamp. Early chondrogenic differentiation of ATDC5 cells for 7 days in vitro, characterized by aggrecan (Acan) and collagen 2a1 (Col2a1) mRNA expression and proteoglycan production, was inhibited by both 50 mM ethanol and 5 mM acetate. Ethanol effects were not blocked by the alcohol dehydrogenase inhibitor 4-methylpyrazole. 50 mM ethanol retarded both ATDC5 cell growth and culture medium acidification. Inhibition of chondrogenic differentiation by 5 mM acetate was associated with elevated phosphorylation of extracellular signal-regulated kinase (ERK)1 and ERK2 and decreased expression of transcription factors Sox9 and Runx2. In acetate-exposed cells, blocking of ERK1 and ERK2 phosphorylation with Trametinib prevented further reduction of Acan and Col2a1 mRNA expression. We conclude that ethanol-derived acetate mediates at least part of the induction of Calcr and Ocstamp expression and that acetate mimics the effects of ethanol on early chondrogenic differentiation.</p>","PeriodicalId":23178,"journal":{"name":"Toxicological Sciences","volume":" ","pages":"181-195"},"PeriodicalIF":4.1,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12448200/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144128481","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Maternal α-cypermethrin and permethrin exert differential effects on fetal growth, placental morphology, and fetal neurodevelopment in mice.","authors":"Benjamin A Elser, Benjamin Hing, Samuel Eliasen, Malik A Afrifa, Naomi Meurice, Farzana Rimi, Michael Chimenti, Laura C Schulz, Michael E Dailey, Katherine N Gibson-Corley, Hanna E Stevens","doi":"10.1093/toxsci/kfaf079","DOIUrl":"10.1093/toxsci/kfaf079","url":null,"abstract":"<p><p>Pyrethroid insecticides represent a broad class of chemicals used widely in agriculture and household applications. Human studies show mixed effects of maternal pyrethroid exposure on fetal growth and neurodevelopment. Assessment of shared pyrethroid metabolites as a biomarker for exposure obscures effects of specific chemicals within this broader class. To better characterize pyrethroid effects on fetal development, we investigated maternal exposure to permethrin, a type I pyrethroid, and α-cypermethrin, a type II pyrethroid, on fetal development in mice. Pregnant CD1 mice were exposed to permethrin (1.5, 15, or 50 mg/kg), α-cypermethrin (0.3, 3, or 10 mg/kg), or corn oil vehicle via oral gavage on gestational days (GDs) 6 to 16. Effects on fetal growth, placental toxicity, and neurodevelopment were evaluated at GD 16. Cypermethrin, but not permethrin, significantly reduced fetal growth and altered placental layer morphology. Placental RNAseq analysis revealed downregulation of genes involved in extracellular matrix remodeling in response to α-cypermethrin. Both pyrethroids induced shifts in fetal dorsal forebrain microglia morphology from ramified to ameboid states; however, the effects of α-cypermethrin were more pronounced. The α-cypermethrin transcriptome of fetal dorsal forebrain implicated altered glutamate receptor signaling, synaptogenesis, and c-AMP signaling. Coregulated gene modules in individual placenta and fetal dorsal forebrain pairs were correlated and overlapped in biological processes characterizing synapses, mitotic cell cycle, and chromatin organization, suggesting placenta-fetal brain shared mechanisms with α-cypermethrin exposure. In summary, maternal exposure to the type II pyrethroid α-cypermethrin, but not type I pyrethroid permethrin, significantly affected placental development, fetal growth, and neurodevelopment, and these effects were linked.</p>","PeriodicalId":23178,"journal":{"name":"Toxicological Sciences","volume":" ","pages":"91-108"},"PeriodicalIF":4.1,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12448229/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144302902","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Detection of developmental toxicity of the anti-COVID-19 drug molnupiravir using gastruloid-based in vitro assays.","authors":"Margaret Carrell Huntsman, Yusuke Marikawa","doi":"10.1093/toxsci/kfaf093","DOIUrl":"10.1093/toxsci/kfaf093","url":null,"abstract":"<p><p>In pharmaceutical drug development, animal tests are traditionally required to conduct comprehensive toxicity assessments before initiating human clinical trials. However, animal tests are time-consuming and can hinder the rapid development of drugs needed to combat urgent health crises, such as the COVID-19 pandemic. Therefore, faster non-animal alternatives are critical to accelerating preclinical toxicity assessments. Molnupiravir, an antiviral medication authorized for emergency use to treat COVID-19, is an oral pro-drug that is metabolized into its active form, N4-hydroxycytidine (NHC). The developmental toxicity of molnupiravir was initially identified in preclinical animal studies. The present study aims to determine whether in vitro assays using gastruloids-three-dimensional aggregates of pluripotent stem cells that mimic axial elongation morphogenesis of early embryos-can effectively detect the developmental toxicity of molnupiravir in a clinically relevant context. In our experiments, NHC at 20 μM significantly impaired the morphological progression and altered the gene expression profiles in gastruloids derived from mouse P19C5 stem cells. Similarly, in a human embryonic stem cell-based morphogenesis model, NHC reduced the aggregate size at 10 μM and induced significant gene expression changes at concentrations as low as 2.5 μM. Notably, these NHC concentrations are comparable to the plasma levels observed in humans (approximately 10.8 μM) following administration of the clinically recommended dose of molnupiravir. These findings demonstrate that gastruloid-based assays can reliably detect the developmental toxicity of NHC at clinically relevant concentrations, supporting their utility as non-animal tools for expediting preclinical developmental toxicity assessments.</p>","PeriodicalId":23178,"journal":{"name":"Toxicological Sciences","volume":" ","pages":"74-90"},"PeriodicalIF":4.1,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12448192/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144683154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The indispensable role of zebrafish as a new approach methodology (NAM) in toxicology.","authors":"Robyn L Tanguay","doi":"10.1093/toxsci/kfaf121","DOIUrl":"https://doi.org/10.1093/toxsci/kfaf121","url":null,"abstract":"","PeriodicalId":23178,"journal":{"name":"Toxicological Sciences","volume":" ","pages":""},"PeriodicalIF":4.1,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144970486","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel computational machine learning pipeline to quantify similarities in 3D protein structures.","authors":"Shreyas U Hirway, Xiao Xu, Fan Fan","doi":"10.1093/toxsci/kfaf007","DOIUrl":"10.1093/toxsci/kfaf007","url":null,"abstract":"<p><p>Animal models are widely used during drug development. The selection of suitable animal model relies on various factors such as target biology, animal resource availability, and legacy species. It is imperative that the selected animal species exhibit the highest resemblance to humans, in terms of target biology as well as the similarity in the target protein. The current practice to address cross-species protein similarity relies on pairwise sequence comparison using protein sequences, instead of the biologically relevant 3D structure of proteins. We developed a novel quantitative machine learning pipeline using 3D structure-based feature data from the Protein Data Bank, nominal data from UNIPROT, and bioactivity data from ChEMBL, all of which were matched for human and animal data. Using the XGBoost regression model, similarity scores between targets were calculated and based on these scores, the best animal species for a target was identified. For real-world application, targets from an alternative source, i.e. AlphaFold, were tested using the model, and the animal species that had the most similar protein to the human counterparts were predicted. These targets were then grouped based on their associated phenotype such that the pipeline could predict an optimal animal species.</p>","PeriodicalId":23178,"journal":{"name":"Toxicological Sciences","volume":" ","pages":"48-56"},"PeriodicalIF":4.1,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143012093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bee venom disrupts vascular homeostasis: apitoxin and melittin trigger vascular cell toxicity and aortic dysfunction in mice.","authors":"Àngel Bistué-Rovira, Montse Solé, Mateu Anguera-Tejedor, Belén Pérez, Laura García-Tercero, Andrea Díaz-Pérez, Zonia Martínez-Benitez, René Delgado-Hernández, Francesc Jiménez-Altayó","doi":"10.1093/toxsci/kfaf086","DOIUrl":"10.1093/toxsci/kfaf086","url":null,"abstract":"<p><p>Bee venom (apitoxin) is a mixture of bioactive molecules, with melittin as its principal component. Although its therapeutic potential is increasingly recognized, its toxic effects on vascular homeostasis remain underexplored. We investigated the impact of apitoxin and melittin on vascular cell viability and mouse aortic function. Cytotoxicity was assessed in cultured endothelial and smooth muscle cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Aortic function was evaluated by mounting thoracic aortas from young male and female C57BL/6J mice in tissue baths. Isometric tension was measured during phenylephrine-induced contractions, as well as endothelium-dependent (acetylcholine) and -independent (sodium nitroprusside) relaxations. To evaluate the roles of nitric oxide (NO) and oxidative stress, we used the NO synthase inhibitor Nω-nitro-L-arginine methyl ester (L-NAME) and the antioxidant superoxide dismutase (SOD), respectively. High-performance liquid chromatography analysis revealed that melittin comprised 43.80% of apitoxin. Both apitoxin and melittin exhibited concentration-dependent cytotoxicity, significantly reducing endothelial cell viability at concentrations ≥5 µg/ml, whereas smooth muscle cells were affected at lower concentrations (≥2.5 µg/ml for apitoxin; ≥1.5 µg/ml for melittin). In functional experiments, apitoxin enhanced phenylephrine-induced contractions at 1 µg/ml and impaired both endothelium-dependent and -independent relaxations at ≥0.1 µg/ml, particularly in males. Although melittin mimicked these effects, higher concentrations (≥5 µg/ml) were required, suggesting that other venom components contribute to the vascular functional toxicity of apitoxin. L-NAME and SOD prevented apitoxin-induced vascular impairments, implicating the NO pathway and oxidative stress. These findings demonstrate that apitoxin impairs vascular cell viability and aortic function at clinically relevant concentrations, underscoring both its vascular risks and therapeutic potential.</p>","PeriodicalId":23178,"journal":{"name":"Toxicological Sciences","volume":" ","pages":"196-207"},"PeriodicalIF":4.1,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144275954","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Estrogen and glucocorticoid receptors co-regulate acrolein-induced respiratory and systemic homeostatic stress responses.","authors":"Devin I Alewel, Katherine M Rentschler, Mette C Schladweiler, Colette N Miller, Stephen H Gavett, Paul A Evansky, Rachel Grindstaff, Wanda C Williams, Urmila P Kodavanti","doi":"10.1093/toxsci/kfaf084","DOIUrl":"10.1093/toxsci/kfaf084","url":null,"abstract":"<p><p>The contribution of neuroendocrine mechanisms of air pollution health effects in females and the extent to which such effects are related to estrogen signaling are unclear. To examine the interactive roles of estrogen (ER) and glucocorticoid receptors (GR) in acrolein-induced respiratory and systemic effects, female Wistar-Kyoto rats were treated daily for 9 days with corn oil (vehicle, 1 ml/kg), fulvestrant (ER-antagonist/degrader, 20 mg/kg), mifepristone (GR antagonist, 30 mg/kg) or fulvestrant + mifepristone, and on days 8 and 9 post-drug-treatment start, rats were exposed nose-only to 0 or 3.2 ppm acrolein for ∼4 h/day. Glucose-tolerance testing was performed following the first exposure. Nasal and lung lavages and blood samples were collected following the second exposure. Fulvestrant and mifepristone pretreatments decreased serum estrogen and progesterone, respectively, and each drug increased adrenocorticotropic hormone in acrolein-exposed rats. Although acrolein-induced nasal and lung protein leakage was reduced in fulvestrant-treated rats, neutrophilic inflammation and pro-inflammatory cytokine increases were exacerbated. However, acrolein-induced airway inflammation was not observed in mifepristone or co-treated rats. Regarding systemic markers of hypothalamic-pituitary-adrenal (HPA) activity, fulvestrant and mifepristone each increased circulating basal leukocytes regardless of exposure, especially total white blood cells and neutrophils. Fulvestrant-induced neutrophilia was slightly dampened in acrolein-exposed females. Fulvestrant also primed multiple adverse acrolein-induced metabolic alterations. Importantly, systemic markers of acrolein-induced HPA activity were not impacted in mifepristone or fulvestrant + mifepristone co-treated rats. These data demonstrate that neuroendocrine co-regulation by ER and GR might explain acrolein susceptibility differences, contributing novel mechanistic information to the growing recognition of gonadal hormone influence in air pollution health effects susceptibility.</p>","PeriodicalId":23178,"journal":{"name":"Toxicological Sciences","volume":" ","pages":"109-125"},"PeriodicalIF":4.1,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12398100/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144259014","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Computationally informed point of departure evaluation for proarrhythmic cardiotoxicity assessment using 3D engineered cardiac microtissues from human iPSC-derived cardiomyocytes.","authors":"Mark C Daley, Peter Bronk, Tae Yun Kim, Arvin H Soepriatna, Cao T Tran, Ulrike Mende, Kareen L K Coulombe, Bum-Rak Choi","doi":"10.1093/toxsci/kfaf094","DOIUrl":"10.1093/toxsci/kfaf094","url":null,"abstract":"<p><p>Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are a promising new approach for in vitro proarrhythmic cardiotoxicity assessment. However, variation due to differentiation batch, individual sample variation, and non-linear responses to test drugs complicate the prediction of proarrhythmic drug concentrations. This study combines a computational human action potential (AP) model of hERG channel block with experimental data from three-dimensional hiPSC-CM engineered microtissues to optimize point of departure (POD) estimation of drug-induced prolongation of AP duration (APD). Computer simulations predicted that APD prolongation from hERG block follows a logistic curve and that >81% hERG block induced early afterdepolarizations (EADs), which significantly shifted the APD response curve. Curve fitting of APD response by logistic, bilinear breakpoint, and maximal curvature was more accurate prior to EAD onset. Goodness-of-fit testing indicated that logistic regression with ≥6 test concentrations was sufficient to accurately estimate PODs. Power analysis, based on experimental variations between batches (n = 14), molds (n = 57), and microtissues (n = 1701), predicted that PODs from 2 ∼ 3 batches with 10 microtissues per mold using a 5% threshold for APD prolongation detected proarrhythmic cardiotoxicity with a negligible false positive rate. We then applied this POD analysis to hiPSC-CM microtissue data after treatment with well-characterized drugs (i.e. cisapride, ranolazine, quinidine, and verapamil). Using bootstrapping, we estimated PODs and confidence intervals that matched concentrations known to cause proarrhythmic effects in patients. This study identified a robust method for calculating PODs for proarrhythmic cardiotoxicity risk in vitro and developed a framework for experimental design in this and other in vitro platforms.</p>","PeriodicalId":23178,"journal":{"name":"Toxicological Sciences","volume":" ","pages":"221-237"},"PeriodicalIF":4.1,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12448244/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144683153","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}